The goal of this project is to identify genes involved in auditory memory and perception in songbirds. Previous work has shown that presentation of conspecific forms a long-lasting memory of that song in a songbird's auditory forebrain area NCM and the establishment of long-term song memory requires mRNA and protein synthesis during several well defined time periods. Little is known about the complex molecular mechanisms underlying song memory. Here, I propose to use a subtractive gene screening method to identify genes regulated in NCM by hearing conspecific song during the first time window, this is 1-3 hours after the onset of song presentation. My preliminary studies have established a conspecific song stimulation paradigm that can effectively induce gene expression in NCM. Two parallel subtractive cDNA libraries will be constructed from the NCM of birds stimulated either with conspecific song or white noise, in which up and down regulated genes are enriched by three cycles of subtractive hybridization. The libraries will be screened with control and induced probes. Candidate genes differentially hybridized with the two probes will be identified, evaluated by Southern Blot analysis and sequenced. Finally, expression of these genes in NCM and their regulation by conspecific song will be confirmed by quantitative in situ hybridization. Identification of these genes is an important first step that will lead to a new area of molecular studies in songbirds field and lead to the eventual understanding of complex molecular mechanisms underlying song learning, memory and perception in songbirds.